Bio-Protocol (Jul 2021)

In vitro Nitrate Reductase Activity Assay of Mycolicibacterium smegmatis Crude Extract

  • Wei Tan,
  • Zhi-Hui Shao,
  • Guo-Ping Zhao

DOI
https://doi.org/10.21769/bioprotoc.4098
Journal volume & issue
Vol. 11, no. 14

Abstract

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Nitrate is one of the major inorganic nitrogen sources for microorganisms. Many bacterial and archaeal lineages can express cytoplasmic assimilatory nitrate reductase (NAS), which catalyzes the rate-limiting reduction of nitrate to nitrite in the nitrate assimilation pathway. Here, we present a detailed protocol for measuring in vitro nitrate reductase (NaR) activity of NAS enzymes from Mycolicibacterium smegmatis crude extract using both physiological and non-physiological electron donors.